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  • 1. Bowers, HA
    et al.
    Brutemark, Andreas
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    F de Carvalho, Wanderson
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Granéli, Edna
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Combining flow cytometry and real-time PCR methodology to demonstrate consumption in Prymnesium parvum2010In: Journal of the American Water Resources Association, ISSN 1093-474X, E-ISSN 1752-1688, Vol. 46, p. 133-143Article in journal (Refereed)
    Abstract [en]

    Harmful algal bloom species can persist in the environment, impacting aquatic life and human health. One of the mechanisms by which some harmful algal bloom species are able to persist is by consumption of organic particles. Methods to demonstrate and measure consumption can yield insight into how populations thrive. Here, we combine flow cytometry and real-time PCR to demonstrate consumption of a cryptophyte species (Rhodomonas sp.) by a toxic mixotrophic haptophyte (Prymnesium parvum). Using flow cytometry, the feeding frequency of a population of P. parvum cells was calculated using the phycoerythrin (PE) fluorescence signal from Rhodomonas sp. and the fluorescence of an acidotropic probe labeling the food vacuoles. Feeding frequency increased in the beginning of the experiment and then began to decline, reaching a maximum of 47.5% of the whole P. parvum population after 212 min. The maximum number of consumed Rhodomonas sp. cells was 0.8 per P. parvum cell, and occurred after 114 min corresponding to an ingestion rate of 0.4 Rhodomonas sp. cells/P. parvum/h. Cells from the feeding P. parvum population were sorted, washed, and subjected to a real-time PCR assay targeting the cryptophyte 18S locus. There was a correlation between cycle threshold (Ct) values and number of consumed prey cells calculated by fluorescence. Overall, this study shows that flow cytometric analysis, of the acidotropic probe and prey pigments, is an efficient and rapid tool in enumerating food vacuoles and the number of prey cells consumed. Furthermore, we suggest that real-time PCR can be applied to cells sorted by flow cytometry, thus allowing for the detection and potential quantification of the targeted prey cells.

  • 2.
    Brutemark, Andreas
    University of Kalmar.
    Contribution of phagotrophy by microalgae to carbon flow in marine food webs2009Doctoral thesis, monograph (Other academic)
  • 3.
    Brutemark, Andreas
    et al.
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Granéli, Edna
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Role of mixotrophy and light for growth and survival of the toxic haptophyte Prymnesium parvum2011In: Harmful Algae, ISSN 1568-9883, E-ISSN 1878-1470, Vol. 10, p. 388-394Article in journal (Refereed)
    Abstract [en]

    Mixotrophy in Prymnesium parvum was investigated using carbon (δ13C) and nitrogen (δ15N) stable isotopes. The experiment was performed in light and dark. In the dark treatment we expected that the mixotrophic P. parvum would rely solely on its prey and therefore reflect the prey isotopic signatures. In the light treatment P. parvum can perform photosynthesis as well as utilize its prey, thus we expect the isotopic signatures to be between the dark mixed cultures and the monocultures, depending on how much prey was utilized. In the light treatment, addition of the ciliate Myrionecta rubra resulted in higher P. parvum cell numbers compared to monocultures. During the experiment, cell numbers in the dark monocultures and the mixed dark cultures did not increase. P. parvum had 2.5-3 times higher cellular phosphorus and nitrogen content in the dark compared to the cultures in the light whereas no difference in carbon content between treatments could be observed. This suggests that P. parvum can utilize nitrogen and phosphorus but not carbon in the dark. It thus seems as if P. parvum relies on photosynthesis to meet the carbon and energy demand required for growth. The expected isotopic signatures “become what you eat…plus a few per mil” were not observed. In the dark treatment, the δ13C did not differ between monocultures and mixed cultures. In the light treatments P. parvum δ13C became less negative then the corresponding dark treatments indicating that P. parvum used CO2 rather than carbon from the added prey. No difference in δ15N between monocultures and mixed cultures could be observed during the experiment. We argue that light is necessary for P. parvum growth and that the ability to utilize nutrients originating from their prey may be important in bloom formation.

  • 4.
    Brutemark, Andreas
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Lindehoff, Elin
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Granéli, Edna
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Granéli, Wilhelm
    Carbon isotope signature variability among cultured microalgae: Influence of species, nutrients and growth2009In: Journal of Experimental Marine Biology and Ecology, ISSN 0022-0981, E-ISSN 1879-1697, Vol. 372, no 1-2, p. 98-105Article in journal (Refereed)
    Abstract [en]

    In this study we have investigated whether the carbon isotopic signature differs between different groups and species of marine phytoplankton depending on growth phase, nutrient conditions and salinity. The 15 investigated algal species, representing the Bacillariophyceae, Chlorophyceae, Cryptophyceae, Cyanophyceae, Dinophyceae and Haptophyceae classes were grown in batch monocultures and analysed for delta(13)C in both exponential and stationary phase. For all the cultured species, delta(13)C signatures ranged from -23.5 parts per thousand (Imantonia sp.) to - 12.3 parts per thousand (Nodulania spumigena) in the exponential phase and from - 18.8 parts per thousand (Amphidinium carterae) to - 8.0 parts per thousand (Anabaena lemmermannii) in the stationary phase. Three species (Dunaliella tertiolecta, Rhodomonas sp.. Heterocapsa triquetra) were also grown under nutrient sufficient and nitrogen or phosphorus deficient conditions. Nitrogen limitation resulted in a more negative delta(13)C signature, whereas no effect could be observed during phosphorus limitation compared to nutrient sufficient conditions. Growth of Prymnesium parvum in two different salinities resulted in a more negative delta(13)C signature in the 26 parts per thousand-media compared to growth in 7 parts per thousand-media. Our results show that the carbon isotopic signature of phytoplankton may be affected by salinity, differ among different phytoplankton species, between exponential and stationary phase, as well as between nutrient treatments.

  • 5.
    Brutemark, Andreas
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Rengefors, Karin
    Anderson, N J
    An experimental investigation of phytoplankton nutrient limitation in two contrasting low arctic lakes2006In: Polar Biology, Vol. 29, no 6, p. 487-494Article in journal (Refereed)
  • 6.
    Carlsson, Per
    et al.
    Lunds Universitet.
    Granéli, Edna
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Granéli, Wilhelm
    Lunds Universitet.
    Gonzalez Rodriguez, Eliane
    IEAPM, Arraial do Cabo, Brazil .
    Carvalho, Wanderson F
    IEAPM, Arraial do Cabo, Brazil .
    Brutemark, Andreas
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Lindehoff, Elin
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Bacterial and phytoplankton nutrient limitation in tropical marine waters, and a coastal lake in Brazil2012In: Journal of Experimental Marine Biology and Ecology, ISSN 0022-0981, E-ISSN 1879-1697, Vol. 418-419, p. 37-45Article in journal (Refereed)
    Abstract [en]

    Bioassay experiments were performed two times (with 2 years in between) in order to investigate if nitrogen(N, ammonium), phosphorus (P, phosphate) and carbon (C, glucose) additions would stimulate the growth ofbacteria and phytoplankton differently in three different tropical aquatic environments. The water and theirindigenous microbial communities were taken from a freshwater coastal lake (Cabiunas), a coastal (Anjos),and an offshore marine station (Sonar) in the Atlantic outside Cabo Frio, Rio de Janeiro State, Brazil. Ammonium,phosphate and glucose were added alone or in combination to triplicate bottles. In the lake, P seemedto be the primary limiting factor during the first experiment, since both bacterial production and phytoplanktongrowth was stimulated by the P addition. Two years later, however, addition of P inhibited phytoplanktongrowth. During both years, C was closely co-limiting for bacteria since CP additions increased the responseconsiderably. For both the coastal and offshore seawater stations, phytoplankton growth was clearly stimulatedby N addition in both years and the bacteria responded either to the P, N or C additions (alone or incombination). To conclude, the results from these tropical aquatic systems show that it is possible that phytoplanktonand bacteria may compete for a common resource (P) in lakes, but can be limited by different inorganicnutrients in marine waters as well as lakes, suggesting that phytoplankton and bacteria do notnecessarily compete for the same growth limiting nutrient in these environments.

  • 7.
    Granéli, Edna
    et al.
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Esplund, Christina
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Lindehoff, Elin
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Brutemark, Andreas
    Novia University of Applied Sciences, Ekenäs, Finland.
    Minimizing economical losses with the help of “real-time” algal surveillance2012In: Eco-Tech 2012 Proceedings, 2012, p. 550-555Conference paper (Other academic)
    Abstract [en]

    Cyanobacterial blooms covering almost the entire Baltic Sea is a yearly feature during July-August. For the tourism industry at Öland island, SE Sweden, the economical losses during the summer 2005 amounted to 17-23 million euros. Remote sensing satellite images show that all the Öland beaches are covered with decomposing algae. In reality, these blooms rarely reach the western side of the island. To more accurately inform the public on the quality of the water for swimming, with the help of volunteers, a daily real-time surveillance of the algal densities on the beaches was performed. The volunteers (from 15 years old to pensioners) were trained at the Linnaeus University, from simple laboratory techniques, to more complicated ones such as identification and enumeration of the toxic cyanobacteria species. By latest 9.00 a.m., the public had access to information on the algal situation on 17 beaches. We could show that: 1) although remote sensing images showed Öland being surrounded by the blooms, our surveillance showed no algal accumulations on the beaches 2) that the real-time warning system boosted public confidence in the local water quality and during the first “Miss Algae”-summer 2006, the economical losses by the tourism industry turned in profits, the gain amounting to 17 million euros, 3) this kind of real-time surveillance is economical feasible due to low-costs involved, but also, the project has a great social value for the volunteers who mostly were pensioners. The volunteers who participated in “Miss Algae” had a good knowledge about the area they monitored (as their houses are located nearby) and could disseminate knowledge to the public in these areas. This kind of project also render a lot of interest regional, national and international, and can be used in advertising campaigns to increase tourism in the areas affected by algal blooms.

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